Date of Award
Master of Science (MS)
College of Science and Mathematics
Thesis Sponsor/Dissertation Chair/Project Chair
Sandra D. Adams
Lee H. Lee
Infection with herpes simplex virus type 1 leads to a lifelong presence of the vims in the host typified by the punctuated recrudescence of herpetic lesions of the orofacial region. Recmdescence of symptoms is due to the ability of the virus to reactivate from a latent state in neurons of the trigeminal ganglia. The latent state characterized by a complete lack of DNA replication, absence of virion stmctural protein production, and the minimal expression of a very limited set of regulatory genes. Reactivation is seen as the return to full viral gene expression, production of new infectious virions, and the emergence of clinical symptoms of herpetic disease. The viral transactivator ICPO has been implicated in both latency and reactivation. PC 12 cells neurally differentiated by nerve growth factor were used as host cells to attempt to generate a quiescent infection. A viral construct deletion mutant GHSV-UL46 that lacks the ICP4 transactivator genes, has a defective US3, and has green fluorescent protein gene attached to gene UL46 was used. Roscovitine, a small molecule inhibitor of cyclin dependent kinases, was used for its ability to prevent ICPO activity via prevention of ICPO’s phosphorylation by cyclindependent kinase 5. Inhibition of ICPO transactivity with roscovitine did not shift expression equilibrium towards latency. The use of an inhibitor of MEK1, PD98059, however, showed better inhibition of GFP expression than either roscovitine or acyclovir. The data indicates that a low-level productive infection was achieved as opposed to a quiescent infection, while studies of small molecule inhibitors posit the possibility that the molecular events of a low-level productive infection differ from those of a fully productive infection.
Caldararo, Mario D., "The Small Molecule Inhibitor Roscovitine Does Not Promote the Establishment of a Quiescent Herpes Simplex Virus Type 1 Mutant GHSV-UL46 Infection in Neurally Differentiated PC 12 Cells" (2012). Theses, Dissertations and Culminating Projects. 789.